JPH09274048A - Pretreatment apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a pretreatment apparatus in which the amount of a cleaning solution required to clean a dilution tank can be reduced and in which a liquid sample or the like can be diffused and mixed in a short time and efficiently by installing a pretreatment tank or the like in which the dilution tank and a drain tank are formed integrally. SOLUTION: A needle 25 sucks a blood sample 24 from a vacuum blood sampling pipe 23. After that, by the motion of a needle holding mechanism 26, it is then moved onto a pretreatment tank 30 in which a dilution tank 30b and a drain tank 30a are formed integrally. In this position, valves 34, 35, 36 are opened, a diluting and cleaning solution 21 is supplied to a needle cleaning block 28, and the outer circumference of the needle 25 is cleaned. Then, the needle 25 is moved onto the dilution tank 30b so as to discharge the blood sample. After that, the valves 34, 35, 36 are changed over, and the solution 21 is discharged and diluted according to a dilution ratio. The diluted sample is mixed in a mixing flow passage 32, and the diluted sample which is returned to the dilution tank 305 is sucked by the needle 25 so as to be taken into a sample loop 33a inside a sample injector 33. At a final stage, the diluted sample which is left is thrown away.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for analyzing a liquid sample in the fields of chemistry and biochemistry, and more specifically, when cleaning a liquid sample using a needle, cleaning of the needle and the sample collected The present invention relates to an apparatus and a method for performing a pretreatment such as diluting.

[0002]

2. Description of the Related Art In glycated hemoglobin analysis using liquid chromatography, for example, a fixed amount of a blood sample in a tube (vacuum blood collection tube) having a rubber cap is collected,
Operations such as analyzing the amount of glycated hemoglobin component in the sample are performed.

In this operation, specifically, a needle-like needle capable of penetrating a rubber cap of a vacuum blood collection tube is accurately penetrated through a pipe passage place having a diameter of about 5 mm at a central portion of the rubber cap, and a predetermined amount of the needle is inserted. It consists of taking a blood sample, diluting the blood sample to any magnification and introducing it into an analytical column. Therefore, pretreatment such as collection (sampling) and dilution is necessary before the sample is actually applied to the column. Further, in the analysis of glycated hemoglobin, a pretreatment of hemolyzing red blood cells before the sample is applied to the analytical column is also necessary. Furthermore,
In order to continuously analyze a plurality of blood samples, a pretreatment such as washing of a needle or the like is required prior to the operation.

As described above, not only in the glycated hemoglobin analyzer, but also in various analyzers, each component of the device is controlled to perform a complicated operation, and special components for washing and dilution are provided. It is equipped.

As an example, an apparatus for analyzing blood, such as a glycated hemoglobin analyzer, will be described below with reference to FIG. In the glycated hemoglobin analyzer, a blood sample can be suctioned with good reproducibility from between vacuum blood collection with a rubber cap, contamination due to bringing in the sample can be eliminated, and reproducibility of about 200 to 400 times dilution is generally good. Is required.

[0006] First, the needle 1 is moved to the sampling position 2, penetrates the rubber cap to the vacuum blood collection tube 3 with the rubber cap, and is lowered, and a predetermined blood sample is sucked by the reciprocating movement of the first syringe 4. Here, in order to aspirate a blood sample with good reproducibility, it is necessary to set the predetermined amount to 3 μl or more. Here, in order to realize the suction by the first syringe 4, the first valve 5 is closed.

Next, the needle is moved to the diluting tank 6 to discharge the sucked blood sample, and a predetermined amount of solution (diluting solution) is ejected from the diluting nozzle 8 by the second syringe 7 to make the blood sample have a predetermined magnification ( It is usually diluted 200 to 400 times).
After the ejection of the solution from the dilution nozzle 8 is completed, the first valve and the second valve 9 are switched to introduce the diluted sample into the sample injector 10 using the 6-way valve, and the diluted sample is supplied to the analytical column. In order to perform the dilution operation with good reproducibility, it is necessary to collect the blood sample in an amount of 3 μl or more.

After the above operation is completed, the diluting tank and the needle are washed in order to analyze a new blood sample. To clean the needle, clean the needle cleaning block 11 with the cleaning pump 1
2, the cleaning solution is discharged from the needle in the needle cleaning tank 13, the third valve 14 is opened, the suction pump 15 is operated, and the contaminated cleaning solution is sucked into the vacuum chamber 16. There is a method of opening the fourth valve 17 at a timing and discharging to the discharging portion 18.

To wash the diluting tank 6, the washing solution is discharged from the needle 1 or the diluting nozzle 8 into the diluting tank until it is full, the fifth valve 19 is opened, and the suction pump 15 is operated to remove the contaminated washing solution. Suck into the vacuum chamber 16,
It is performed by opening the fourth valve 17 at a certain timing and causing the discharging portion 18 to discharge the gas.

Here, in order to reduce the contamination between the samples, the needle 1, the needle washing tank 13,
Needle washing block 11, dilution tank 6 and sample supply unit 1
It is required to clean the inside of the pipe connecting 0 and these.

[0011]

SUMMARY OF THE INVENTION In the conventional glycated hemoglobin analyzers and the like, particularly conventional analyzers that handle liquid samples such as blood samples, in order to improve reproducibility, a blood sample from a vacuum blood collection tube is used in an amount of 3 micron. Aspirate more than 1 and the dilution ratio of general blood sample is 200 to 400
Since it is double, the capacity of the dilution tank is 1500 μl.
As a result, it is necessary to make the washing operation 3 or more.
There is a problem that a large amount of cleaning solution of about 000 μl or more is required.

Further, in the conventional apparatus, a dedicated suction pump and a dedicated valve are required to wash the dilution tank, and a dedicated liquid feed pump and a dedicated valve are also required to wash the needle. Therefore, there is a problem that not only the apparatus becomes complicated, but also the operation of each component becomes complicated, and as a result, a long time is required for the pretreatment.

Further, in the conventional device as shown in FIG. 1,
After the diluting tank 6 is filled with the cleaning solution discharged from the needle 1 or the diluting nozzle 8, the valve 19 is opened and suction is performed by the suction pump 15, so that the upper end of the diluting tank is insufficiently cleaned, and contamination is caused. Not only is it possible to eliminate the possibility that this will occur, but it may cause problems such as clogging of the needle and valve in some cases. Therefore, periodically diluting the diluting tank 6, valve 19 and suction pump 15 manually. Such as having to wash
There is a problem that complicated maintenance is required.

In particular, during vacuum blood collection with a rubber cap, since the needle penetrates the rubber cap, there is a possibility that wear debris of the rubber cap adhering to the needle may flow out to each portion, and in this case, it reaches the analytical column. There is a problem that clogging may occur in all routes.

Further, after suctioning the liquid sample, the needle moves to the diluting tank, and after ejecting the liquid sample into the diluting tank, the diluting solution is ejected from the diluting nozzle so as to have an arbitrary magnification (200 to 400 times). At this time, the diluting solution is jetted into the diluting tank at an angle from an obliquely upward direction deviating from the center of the diluting tank and swirling is generated in the diluting tank to improve the diluting efficiency. However, since the amount of the diluted solution to be ejected is determined by the dilution ratio, there is a problem that the blood sample and the diluted solution are insufficiently mixed especially when the dilution ratio is low.

Further, particularly when blood is used as a liquid sample, the pretreatment thereof may include not only dilution but hemolysis of blood cells, but hemolysis tends to be insufficient only by the above-mentioned dilution.

[0017]

The present inventor has completed the present invention as a result of extensive studies to solve the problems found in the prior art. That is, the present invention, at least, a needle for collecting a fixed amount of a liquid sample to be pretreated, a needle holding mechanism for holding the needle so as to be movable up and down, and a needle holding mechanism for holding the needle movable in a horizontal direction. The base, the needle wash block installed in the needle holding mechanism that can move horizontally with the needle but does not move up and down, the dilution tank for diluting the liquid sample, and the drain tank for discarding the drainage liquid. A pretreatment tank integrally formed,
The pretreatment device comprises a mixing flow path for mixing a diluted liquid sample, a pipe connecting these components, and at least two syringes. Hereinafter, the device of the present invention applied as a pretreatment unit of a glycated hemoglobin analyzer will be described in detail with reference to the drawings.

FIG. 2 is a diagram showing the configuration of the present invention. The pretreatment device according to the present invention performs collection of liquid sample, dilution, mixing, and cleaning of each component to prevent contamination as pretreatment. In order to perform such preprocessing,
The present invention uses a diluting solution for diluting a liquid sample and a washing solution for washing each part of the device. As these solutions, it is possible to use two or more different solutions, but it is also possible to use a single solution 21 as shown in FIG. That is, in the glycated hemoglobin analyzer, it is possible to perform dilution and washing with, for example, a single solution having a salt concentration capable of hemolyzing erythrocytes and containing a detergent for washing. In the device shown in FIG.
The diluting / washing solution 21 not only dilutes the blood sample 24 in the vacuum blood collection tube 23 having the rubber cap 22 but also has a function as a washing solution for the needle 25 and the like after the blood sample is taken. When a low salt concentration solution is used as the diluting / washing solution, hemolysis is likely to be incomplete by simply diluting the blood sample, but sufficient hemolysis can be achieved by sufficient mixing in the mixing channel described later. It can also be realized.

In the apparatus of FIG. 2, a first syringe 31, a tank for holding the washing / diluting solution 21, a first valve 34 and a flow path (solution suction system) composed of a pipe connecting these, a first syringe 31. , A sample injector 33 using a normal 6-way injection valve, a needle 25, a flow path (sampling system) composed of a first valve 34 and a second valve 35, and a pipe connecting these, a first syringe 31, a needle wash A flow path (needle cleaning system) composed of the block 28, the first valve 34, the second valve 35, the third valve 36, and a pipe connecting these
A flow path (mixing flow path system) and a blood sample that are composed of the first syringe 31, the mixing flow path 32, the diluting tank 30b, the first valve 34, the second valve 35, the third valve 36, and the pipes connecting these It includes a system including a second syringe 29 for suctioning, a needle 25, and a pipe connecting these. As described above, the configuration of the present invention shown in FIG. 2 has the first syringe 31 for sucking and discharging the diluting / washing solution and the second syringe 29 for sucking and discharging the blood sample. It is also possible to load a syringe added separately.

In the apparatus of the present invention shown in FIG. 2, prior to the actual pretreatment of the liquid sample described later, the syringe is moved in advance and each valve is switched to fill each system with the diluting / washing solution. Keep it. However, for the mixing flow channel system, a dilution tank 3 is used by using a first syringe.
The dilution / washing solution filled with 0b is moved back to between the mixing channel and the diluting tank, and the air layer is positioned on the diluting tank 30b side so that the diluted sample and the washing / diluting solution can be positioned with the air layer in between. It is preferable.

In the example shown in FIG. 2, since the needle 25 penetrates the rubber cap 22 of the vacuum blood collection tube 23, it has a needle shape, but there is no particular limitation on its length or specific shape.

The needle 25 is held by a needle holding mechanism 26 so as to be vertically movable in the vertical direction shown by A in the figure. The needle holding mechanism 26 is held by a base (not shown) so as to be movable in the horizontal direction shown by B in the figure. A needle washing block 28 is attached to the needle holding mechanism 26. That is, the needle 25 is the cleaning block 28.
Although it is configured so as to be able to move up and down by passing through, it does not move up and down, so that the relative positional relationship with the needle washing block 28 is changed by the up and down movement of the needle 25.

First, the needle 25 is moved in the horizontal direction (B
Movement of the blood sample 24 from the vacuum blood collection tube 23
Move to the top of the position for collecting. At this position, the needle holding mechanism 26 descends downward (direction A), and the needle 25 penetrates the rubber cap 22 and is immersed in the blood sample 24 (state of 25a). By moving the second syringe 29 in this state, a predetermined amount of blood sample is sucked. The amount of aspiration in glycated hemoglobin analysis is usually 3
It is about a microliter.

After the blood sample is sucked, the needle 25 cleans the outer circumference of the needle by the movement of the needle holding mechanism 26 and the base, so that the diluting tank 30b for diluting the liquid sample is obtained.
And the drainage tank 30a for discarding the drainage moves to the drainage tank 30a of the pretreatment tank 30 integrally formed. At this position, the valves 34, 35 and 36 on the path from the first syringe 31 to the needle washing block 28 are opened, the needle washing system is opened to supply the dilution / washing solution 21 to the needle washing block 28, and the outer circumference of the needle 25. To wash. The needle washing block 28 allows the needle to penetrate therethrough,
Further, it is a block in which a columnar hollow portion having an inner diameter slightly larger than the outer diameter of the needle, which serves as a cleaning passage for the needle, is formed in the vertical direction. This hollow portion communicates with a cleaning solution introducing passage provided on the upper side surface of the block. The washing solution introducing passage is connected to the first syringe 31 via a valve, and when the needle washing system is opened, the diluting / washing solution 21 is guided from the introducing passage to the needle washing passage and propagates along the outer periphery of the needle 25. The outer periphery is cleaned and discarded into the drainage tank 30a. According to such a configuration, it is possible to simply discharge the diluted / cleaning solution contaminated by gravity without using a special suction pump or the like for discharging the liquid. Of course,
The drainage tank 30a can be connected to a pump or the like via a vacuum chamber or the like.

The needle washing passage is a diluting / washing solution passage for washing the needle, and at the same time holds the needle 25 penetrating the needle washing block 28,
Since it also serves as a guide when the needle 25 moves up and down, its length is preferably as long as possible. However, as will be described later, since the needle 25 is moved up and down when cleaning the needle, it can be appropriately determined in consideration of the range of this up and down movement.

The upper end of the needle washing passage formed vertically in the needle washing block 28 is sealed by a seal member such as an O-ring so that when a diluting / washing solution is supplied, it does not overflow from the upper portion and the lower end. The liquid is drained only from the open end. The seal member such as the O-ring does not suppress the vertical movement of the needle 25. By moving the needle up and down in this state, the effect of wiping off the abrasion residue and the like of the rubber cap attached to the outer periphery of the needle 25 can be obtained. Have. It is advisable to perform such vertical movements about three times. The width of the movement is particularly preferably such that the entire needle comes into contact with the O-ring or the like, but it is sufficient if at least the portion penetrating the rubber cap comes into contact. Further, in order to improve the transmission of the diluting / washing solution to the outer circumference of the needle, the diluting / washing solution introducing passage is configured to intersect the needle washing passage at a right angle, and
The inner diameter of the diluting / washing solution introducing passage is preferably smaller than the inner diameter of the needle washing passage.

As described above, the drainage tank 3 of the pretreatment tank 30 in which the dilution tank for diluting the liquid sample and the drainage tank are integrally formed.
The needle 25 whose outer periphery has been cleaned by moving it up and down a plurality of times while flowing the diluting / cleaning solution on 0a moves horizontally on the diluting tank 30b of the pretreatment tank 30 and descends to the bottom thereof. At this position, the system from the second syringe 29 to the needle 25 is opened, and the whole amount of the blood sample sucked into the needle is discharged. After that, the valve is switched to open the sampling system from the first syringe 31 to the needle 25,
The dilution / washing solution 21 is slowly discharged according to the dilution ratio. At this time, the capacity of the diluting tank 30b may be less than or equal to the capacity of the diluted sample that appears by dilution as long as it is equal to or more than the amount of the sample to be actually used for the analysis. Dilution tank 30b and drainage tank 3
0a is formed integrally, but its edge 30c
Is so low that the overflowed diluted sample is spilled into the drainage tank 30a. The drainage tank 3
The diluted sample overflowing to 0a can be discharged only by gravity.

As can be seen from the figure, this diluting tank 30b is constructed so that its horizontal cross-sectional area increases from the bottom to the top. The shape of the diluting tank 30b may be a taper shape (tapered cone shape) so that the change of the cross-sectional area is continuous, but as shown in the figure, changes in about three stages (three different cross-sectional areas are different). It may be a step change (like combining cylinders). With such a configuration, the diffusion of the blood sample into the diluting / washing solution is insufficient and a concentration gradient occurs in the sample after dilution, and a relatively high concentration portion overflows into the drainage tank 30a and is lost. You can prevent problems. Also,
Since the sample after dilution is sucked again by the needle 25 as described below, it is preferable to consider the inner diameter of the bottom so that the tip of the needle 25 can reach the bottom.

According to the knowledge of the present inventor, the bottom portion of the diluting tank 30b has a cross-sectional area of 0.
The circular shape is about 5 mm to 1.5 mm larger, and the tapered shape as shown in FIG. 3 is particularly preferable. Also,
The vicinity of the upper end portion is preferably about 4 to 6 times the cross-sectional area of the bottom portion in order to improve the diffusion state during dilution.

Next, the valves 34, 35 and 36 are switched to switch the first syringe 31 to the dilution tank 30b of the pretreatment tank 30.
The mixing flow channel system reaching the lower end of the column is opened, and the first syringe 31 is reciprocated to reciprocate the diluted sample in the flow channel indicated by C in the figure. Here, as is clear from the figure, the pipe in the vicinity of the diluting tank 30b of the mixing flow channel system is a special mixing flow channel 32 in which a large portion and a small portion of the cross section of the pipe are continuous at a plurality of locations. The mixing channel 32 is effective for generating an appropriate turbulent flow by changing the flow rate of the diluted sample passing through the portion, improving the mixing efficiency, and further for hemolyzing the blood sample. The shape of the mixing flow path is not limited to that shown in the figure, and there is no limitation as long as the cross-sectional area changes so that the turbulent flow can be generated by changing the flow velocity of the passing liquid.
The path is preferably configured to have the same capacity as the diluting tank 30b or have a volume larger than that. The diluted sample mixed in the mixing channel system is finally returned to the dilution tank. In addition, in the mixing channel system, dilution is performed with an air layer in between.
The sample is located after washing solution and dilution. During this operation using the mixing channel system, the needle is preferably located in the diluting tank 30b, but may be once moved to the upper portion of the diluting tank 30b or the like.

After mixing the diluted sample in the mixing flow path system, the sampling system from the first syringe 31 to the needle 25 is opened for the whole amount of the diluted sample returned to the dilution tank 30b.
The first syringe 31 is moved and sucked. As is clear from the figure, a general sample injector 33 using a 6-way injection valve is arranged in the sampling system from the first syringe 31 to the needle 25, and the diluted sample aspirated from the needle 25 A fixed amount is taken into the sample loop 33a in the sample injector 33. Here, when the flow path capacity from the needle 25 to the sample loop 33a is larger than the capacity of the diluting tank 30b, the needle 25 is once moved to the upper part of the diluting tank 30b in order to take in air following the diluted sample. May be. In particular, it is preferable that the volume of the flow path leading to the sample loop 33a be approximately the same as the volume of the diluting tank 30b. More preferably, the flow path capacity up to the valve 35 is set to be approximately the same as the capacity of the diluting tank 30b, and the sample loop 33a is positioned in the middle in terms of capacity.

After diluting the whole amount of the sample with the needle 25, the mixing channel system is opened with the tip of the needle positioned at the bottom of the diluting tank, and the first syringe is moved to discharge the diluting / washing solution. , Wash the dilution tank and the mixing channel. By this washing, the diluted sample attached to the outer circumference of the needle is also washed. The discharge amount of the dilution / cleaning solution is the dilution tank 30b.
The volume is set to exceed the capacity of the above, and it is operated so as to overflow into the drainage tank 30a. After this, move the first syringe to move the diluted / washed solution remaining in the diluted layer back up between the mixing channel and the dilution tank, and in the next pretreatment operation, sandwich the air layer and dilute the sample and wash / dilute. Allow the solution to be located.

As the final stage of the pretreatment operation, the needle 2
5 is moved from the diluting tank 30b to the draining tank 30a, the sampling system is opened, the diluting / cleaning liquid is discharged by the movement of the first syringe 31, and the diluted sample remaining in the sampling system is discarded. This completes the cleaning of the sampling system including the inside of the needle. Therefore, the sample after dilution remaining in the sampling system during the next sampling operation will not cause contamination.

The diluted sample taken into the sample loop 33a is then provided to a glycated hemoglobin analysis column packed with a cation exchange resin (not shown).

[0035]

In the pretreatment device of the present invention, when the diluted sample obtained by diluting a liquid sample such as blood exceeds the capacity of the diluting tank, the portion exceeding this capacity overflows into the draining tank and is discarded. To be done. In the usual glycated hemoglobin analysis, it is necessary to dilute the blood sample by 200 to 400 times, but on the other hand, the amount of the blood sample collected by the needle cannot be 3 μl or less due to the problem of reproducibility, As a result, if it is attempted to hold all of the sample after dilution, the capacity of the diluting tank becomes large, and naturally, a large amount of washing solution is required for washing the diluting tank, and further, the diffusion and mixing of the blood sample in the diluting tank are insufficient. It is easy to become. On the other hand, in the present invention, the capacity of the diluting tank is positively reduced, and the diluted sample that overflows from the diluting tank is used for the primary cleaning of the diluting tank.

By thus reducing the capacity of the diluting tank, the amount of the cleaning solution required for cleaning the diluting tank can be reduced in the present invention. Moreover, in the present invention, by devising the shape of the diluting tank and further equipping the diluting tank with the mixing channel, the liquid sample and the like in the diluting tank can be diffused and mixed efficiently in a short time. With such a configuration, particularly in a pretreatment device of a glycated hemoglobin analyzer that handles a blood sample as a liquid sample, efficient diffusion and mixing are possible, but in this case, sufficient hemolysis of the blood sample is also possible. There is an effect that it can be realized. In addition, it is possible to easily obtain the mixing degree according to the type of the liquid sample by changing the number of times of movement of the syringe and adjusting the number of times of reciprocating the mixing flow path.

In the present invention, as shown in the drawing, a single syringe and three valves for switching the system can be used for pretreatment such as washing of the needle, the diluting tank and the mixing flow path. It is possible to simplify and shorten the time required for pretreatment.

Further, according to the present invention, it is possible to clean the columnar hole serving as the diluting tank by discharging the cleaning solution from the syringe, and the cleaning solution discharged in excess of the capacity of the diluting tank is adjacent to the cleaning solution. Since it overflows into the drainage tank, it is effective in reducing contamination due to contamination of the upper end of the dilution tank.In addition, conventional equipment such as a suction pump dedicated to cleaning the dilution tank, a vacuum chamber and a dedicated valve can be used. No need to use used parts. Further, since the cleaning solution overflows from the diluting tank, dirt does not adhere to the upper end portion of the diluting tank, and as a result, the diluting tank can be maintenance-free.

In the present invention, the cleaning of the needle for collecting a fixed amount of a liquid sample such as blood is sufficient, and in particular, the wear of the rubber cap attached to the needle by the sealing member such as the O-ring attached to the needle cleaning block. Since the cleaning liquid is attached to the needle and the outer periphery of the needle is cleaned while wiping off the dust, the abrasion dust of the rubber cap may be discarded in the drainage tank and may flow out to the piping in the device, causing clogging. Also has the effect of being small.

[0040]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the case where the present invention is applied to a pretreatment device for a glycohemoglobin analyzer will be described based on Examples, but the present invention is not limited to these Examples.

As a pretreatment device for a glycohemoglobin analyzer, generally, the time required for the pretreatment is 120 seconds or less, and it is brought in from a preblood sample (contamination).
Is less than 1%, dilution ratio is about 250 times, its reproducibility (C
V) is required to be 10% or less, and not only dilution but also hemolysis of red blood cells is required.

The structure of the entire pretreatment apparatus used is as shown in FIG. As the diluting / washing solution 21, a commercially available hemolytic agent for glycated hemoglobin analysis (hemolytic agent C, Tosoh Corporation)
As a blood sample 24, blood was collected in a vacuum blood collection tube 23 with a rubber cap 22 and EDTA added as a blood coagulation inhibitor, and after 3 minutes of centrifugation at 3000 rpm for 10 minutes, the viscosity was increased. I used the one that was higher.

The first syringe 31 for sucking and discharging the solution has a capacity of 2500 microliters, and the second syringe 29 for sucking and discharging the sample has a capacity of 2 microliters.
A 50 microliter syringe, commercially available 3-way solenoid valves (made by Advance Co., Ltd.) for the first valve 34, the second valve 35, and the third valve 36, respectively, and the sample injector 3
A commercially available 6-way injection valve (manufactured by Rheodyne Co., Ltd.) was used as 3. The capacity of the sample loop 33a of this sample injector is 10 μl.

The needle 25 has an outer diameter of 1.25 mm.
The length was 15.5 cm and the hole shape was 0.6 mm and the opening was opened 4.2 mm from the tip of the needle. The mixing flow path connected to the diluting tank 33b was made of a Teflon tube having an inner diameter of 2 mm and a length of 20 cm, and was processed so as to have an uneven cross-sectional area by squeezing a part thereof so as to generate turbulent flow. I used one.

A PMMA (acrylic resin) tank was used as the pretreatment tank 30 in which a diluting tank for diluting the liquid sample and a drainage tank were integrally formed. A general view of the pretreatment tank is shown in FIG. 3a, and details of the diluting tank 30b are shown in FIG. 3b.

The area of the opening of the drainage tank 30a is 112.2.
6 mm 2, and the axis of the hole of the diluting tank and the axis of the hole of the drainage tank are separated by 14 mm. The total length (a) of the diluting tank 30b is 47 mm, and the inner diameter is 2.5 from the bottom to the point 7 mm.
A cylindrical hole having an inner diameter of 3 mm, a bottom surface of 7 mm to 43 mm, and an inner diameter of 3 mm, a bottom surface of 43 mm to 47 mm, and an inner diameter of 5 mm. . The content volume is 376.18 microliters, but the volume with the needle lowered to the bottom of the diluting tank is about 310 when the volume for the needle is subtracted.
It becomes microl. The height (b) around the dilution tank is 4.5 m
However, since the limit to the drainage tank is the same height as the upper end of the diluting tank and the width (c) is 1 mm, the diluted sample etc. overflowing from the diluting tank flows into the draining tank and is discharged from the pretreatment tank. It has a structure that does not leak to the outside.

The needle washing block 28 has a total length of 40 m.
A block made of PMMA (acrylic resin) having a size of m, a width of 20 mm, and a depth of 40 mm was used as a needle washing passage having a cylindrical hole having an inner diameter of 1.8 mm and a total length of 40 mm. An inner diameter of 0.8 mm that connects to the needle washing path at a position 5 mm from the upper end of the needle washing path at a right angle.
Of the washing liquid introduction path, and an O-ring having an inner diameter of 1.2 mm is fixed as a sealing material at the opening at the upper end of the needle washing path, and a hollow portion having an inner diameter of 1.8 mm and a length of 5 mm is further provided at the lower end. The columnar nozzle is fixed so that the cleaning liquid can be easily discarded in the drainage tank 30a.

The pipes other than the mixing channel 32 have an inner diameter of 0.
An 8 mm Teflon tube was used.

The amount of the blood sample collected by the needle was set to 3 μl, and the washing / diluting solution discharged to the diluting tank was 600
The discharge rate was 131 μl / sec. The capacity of the diluting tank is 310 μl, and 290 μl overflows into the drainage tank 30a. The needle is washed with 900 μl of diluting / washing liquid while reciprocating the needle three times up and down, and the diluting tank and the mixing channel are washed with a total volume of 36 parts.
While the amount was 7.18 μl, 1342 μl of the diluted / washing solution was discharged.

The operation of mixing the diluted sample in the mixing channel is as follows.
The sample was diluted by reciprocating the first syringe, and the sample was reciprocated three times in the flow path. After the pretreated blood sample is applied to the column filled with cation exchange resin by the sample injector, the excess diluted sample is discharged from the needle into the drainage tank, and further 300 μl of diluted / washed solution is discharged. Then, the piping of the sampling system and the needle were washed.

The time required for the above operation was about 110 seconds. Further, the pretreatment operation by the pretreatment apparatus of the present invention, especially the mixing operation was sufficient, and as a result, the hemolysis of blood cells was also sufficient, so the analysis result was good. After performing this pretreatment operation, a similar pretreatment operation was performed using a liquid sample that did not contain glycated hemoglobin components.As a result, the chromatogram peak that was probably due to the remaining blood sample (contamination) was It was hardly recognized. Further, the area of the obtained chromatogram showing the absolute amount of blood cells was almost the same as that in the case where the manual pretreatment (dilution) of 250 times was performed.

When the reproducibility of the pretreatment (dilution) was observed using the same blood sample, the reproducibility after 10 pretreatment operations was cv = 1.26%.

Using the glycohemoglobin analyzer to which the pretreatment device of the present invention described above was applied, 8000 samples of human blood collected in a vacuum blood collection tube were analyzed, and as a result, adhered to the needle in the drainage tank. The scraps of the rubber cap were discarded, no contamination (dirt) was confirmed in the diluting tank, and no trouble such as clogging occurred.

[Brief description of drawings]

FIG. 1 is a diagram showing a conventional pretreatment device applied to a glycated hemoglobin analyzer.

FIG. 2 is a diagram showing an analyzer of the present invention applied to a glycated hemoglobin analyzer.

FIG. 3 is a diagram showing details of a pretreatment tank in which a diluting tank for diluting a liquid sample used in an example and a drainage tank are integrally formed. In the figure, FIG. 3a shows the whole pretreatment tank, and FIG. 3b shows only the diluting tank.

[Explanation of symbols]

 1 Needle 2 Sampling Position 3 Vacuum Blood Collection Tube with Rubber Cap 4 First Syringe 5 First Valve 6 Diluting Tank 7 Second Syringe 8 Diluting Nozzle 9 Second Valve 10 Sample Injector 11 Needle Washing Block 12 Washing Pump 13 Needle Washing Tank 14th Three valves 15 Suction pump 16 Vacuum chamber 17 Fourth valve 18 Discharge part 19 Fifth valve 21 Dilution / washing solution 22 Rubber cap 23 Vacuum blood collection tube 24 Blood sample 25 Needle 26 Needle holding mechanism 28 Needle washing block 29 Second syringe 30 Dilution Pretreatment tank in which a tank and a drainage tank are integrally formed 30a Drainage tank 30b Dilution tank 30c Threshold between drainage tank and dilution tank 31 First syringe 32 Mixing channel 33 Sample injector 33a Sample loop 34 First valve 35 Second valve 36 Third valve

Claims (3)

[Claims] 1. A needle for collecting at least a fixed amount of a liquid sample to be pretreated, a needle holding mechanism for holding the needle so as to be vertically movable, and a needle holding mechanism for holding the needle so as to be movable in a horizontal direction. The base, the needle wash block installed in the needle holding mechanism that can move horizontally with the needle but does not move up and down, the dilution tank for diluting the liquid sample, and the drain tank for discarding the drainage liquid. A pretreatment device comprising a pretreatment tank integrally formed, a mixing flow path for mixing a diluted liquid sample, a pipe connecting these components, and at least two syringes. 2. The pretreatment apparatus according to claim 1, wherein the liquid sample is a blood sample. 3. The pretreatment device according to claim 1, which is applied as a pretreatment part of a glycated hemoglobin analyzer.